fluorescein isothiocyanate (fitc)-5-bromo-2′-deoxyuridine (brdu) cell proliferation detection kit  (Keygen Biotech)

Journal: Clinics

Article Title: microRNA-181a-5p impedes the proliferation, migration, and invasion of retinoblastoma cells by targeting the NRAS proto-oncogene

doi: 10.1016/j.clinsp.2022.100026

Figure Lengend Snippet: MiR-181a-5p inhibits RB cell proliferation and metastasis and induces cell apoptosis. (A) qRT-PCR showed that miR-181a-5p was upregulated in Y79 cells transfected with miR-181a-5p mimics and downregulated in SO-RB50 cells transfected with miR-181a-5p inhibitors. (B-D) Cell Counting Kit 8 (CCK-8) and 5′-bromo-2′-deoxyuridine (BrdU) assays showed that miR-181a-5p mimics inhibited the cell proliferation, while miR-181a-5p inhibitors promoted the cell proliferation. (E-F) Transwell assay showed that miR-181a-5p mimics inhibited the migration and invasion of Y79 cells, while miR-181a-5p inhibitors promoted the migration and invasion of SO-RB50 cells (magnification: × 200; bar: 25 μm). (G) Flow cytometry showed that miR-181a-5p mimics promoted apoptosis in Y79 cells, while miR-181a-5p inhibitors inhibited the apoptosis of SO-RB50 cells. Mimics NC indicates the negative control of miR-181a-5p mimics, and inhibitor NC indicates the negative control of miR-181a-5p inhibitors. The data are shown as the mean±standard deviation from three independent experiments. **p < 0.01 and ***p < 0.001.

Article Snippet: A BrdU cell proliferation detection kit (Abcam, Shanghai, China) was used to detect the RB cell proliferation.

Techniques: Quantitative RT-PCR, Transfection, Cell Counting, CCK-8 Assay, Transwell Assay, Migration, Flow Cytometry, Negative Control, Standard Deviation

Journal: Clinics

Article Title: microRNA-181a-5p impedes the proliferation, migration, and invasion of retinoblastoma cells by targeting the NRAS proto-oncogene

doi: 10.1016/j.clinsp.2022.100026

Figure Lengend Snippet: miR-181a-5p exerts an anti-tumor effect in RB by regulating NRAS. (A-C) qRT-PCR and western blotting assays showed that the expression levels of NRAS mRNA and protein were upregulated in the miR-181a-5p mimics+pcDNA-NRAS group compared with the miR-181a-5p mimics group and downregulated in the miR-181a-5p inhibtors+si-NRAS group compared with the miR-181a-5p inhibitor group, while miR-181a-5p expression showed no significant difference. (D-F) CCK-8 and BrdU assays showed that cell proliferation was promoted in the miR-181a-5p mimics+pcDNA-NRAS group compared with the miR-181a-5p mimics group and inhibited in the miR-181a-5p inhibitors+si-NRAS group compared with the miR-181a-5p inhibitor group. (G-H) Transwell assay showed that the migration and invasion of RB cells in the miR-181a-5p mimics+pcDNA-NRAS group was promoted compared with the miR-181a-5p mimics group and inhibited in the miR-181a-5p inhibitors+si-NRAS group compared with the miR-181a-5p inhibitor group (magnification: × 200; bar: 25 μm). (I) Flow cytometry showed that the apoptosis of RB cells in the miR-181a-5p mimics+pcDNA-NRAS group was inhibited compared with the miR-181a-5p mimics group and promoted in the miR-181a-5p inhibitors+si-NRAS group compared with the miR-181a-5p inhibitor group. Protein bands were quantified by comparison with the endogenous control GAPDH. The data are shown as the mean±standard deviation from three independent experiments. ** p < 0.01 and *** p < 0.001.

Article Snippet: A BrdU cell proliferation detection kit (Abcam, Shanghai, China) was used to detect the RB cell proliferation.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, Transwell Assay, Migration, Flow Cytometry, Standard Deviation